Oligosaccharide-producing lactic acid bacteria were isolated from young radish and identified as Weissella cibaria YRK005. Oligosaccharides from W. cibaria YRK005 were purified using size-exclusion high-performance liquid chromatography, and their structure was determined. The oligosaccharides consisted of glucose only, 1 H-nuclear magnetic resonance analysis revealed that the oligosaccharides consisted of 93.8% α-1,6 glycosidic linkages and 6.2% α-1,4 glycosidic linkages, and molecular weight was determined to be 1.1210 2 Da. When the oligosaccharides' prebiotic effect on probiotic strains was examined, the oligosaccharides promoted the growth of four probiotic strains, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus pentosus, compared to fructo-oligosaccharides as a reference. These results indicated that oligosaccharides from W. cibaria YRK005 could be used as potential prebiotics.

The glycoprotein hormone family consists of follicle-stimulating hormone (FSH; GTH1), luteinizing hormone (LH; GTH2), and thyroid-stimulating hormone (TSH), which are secreted by the pituitary gland in all mammalian species, and chorionic gonadotropin, which is secreted by placental trophoblast cells in primates and equids. These hormones consist of non-covalently associated α-, β- subunits. Within a species, the amino acid sequence of α-subunit is identical across all glycoprotein hormones and is encoded by a single gene. The αβ dimer is the active form of the hormone, and biological specificity is conferred by the β-subunit. Both of α and β subunit of eel FSH has two N-glycosylation sites (α-subunit: Asn56 and Asn79; β -subunit: Asn5 and Asn22, respectively). In the present study, we constructed deglycosylated mutants at single and double sites in each subunits of eel FSH for identification of Asn linked oligosaccharides' biological role. Mutant cDANs were cloned into pcDNA3 expression vector and transiently transfected into CHO suspension cells. The quantity of rec-eelFSHs were quantified by sandwich ELISA system, using monoclonal antibodies produced in our lab. The wild type rec-FSH protein was detected at the predicted molecular weight of 34 kDa by western blot. The molecular weight of deglycosylated mutants at single site decreased with about 4 kDa and of mutants at double sites decreased with 8 kDa. After PNGase treatment in the rec-eel FSH proteins, molecular weight also decreased to 7-8 kDa. We generated stably parental cell lines, engineered to express a β-arrestin 2EA fusion protein, expressing eel FSHR and C-terminal deleted mutant. 2 out of 5 receptor cells each were selected by G-418 and we tested these cell lines in a receptor functionality using PathHunter β arrestin assay (DiscoverX). Follicle stimulating hormone acts through binding to its specific receptor. Binding of ligand to the receptor activates the adenosine 3',5'-cyclic monophosphate (cAMP) pathway (McFarland et al., 1989; Ji and Ji, 1991a; Rose, 1998) and the inositol 1phosphate (IP1) the second messenger systems. After stimulation of eelFSH receptor stably transfected Parental CHO cells with FSH wild type and mutant hormones as a ligand, production of cAMP and IP-1 were evaluated (Cisbio). cAMP IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79; and βΔ5 were 33.1; 1154.7; 22; 410 and 311.9 ng/ml, respectively. IP-1 IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79 and βΔ5 were 6.8; 7.1; 4.4; 3.8 and 10.2 ng/ml, respectively too. The cAMP activation was greatly decreased in the αΔ56αmutant. Thus, the site of α56 oligosaccharide in the eel plays an pivotal role for the cAMP stimulation using eel FSH receptor cell lines. In the IP-one assay, the activity in the αΔ56 and βΔ5 mutants was a little decreased than the wt. The biological roles of N-linked oligosaccharides in GPCR internalization are going to be estimated by measuring β arrestin recruitment system.

The glycoprotein hormone family consists of luteinizing hormone (LH), follicle stimulating hormone (FSH) and thyroid stimulating hormone, which are secreted by the pituitary gland in all mammalian species, and choriogonadotropin (CG), which is secreted by the placenta in primates and equids. The hormones are composed of a common α subunit and a hormone specific β subunit which are non-covalently associated. Recent advances in biotechnology, particularly in the production of recombinant proteins, have provided opportunities to produce sufficient quantities of recombinant fish GTHs using various expression hosts. Japanese eel Anguilla japonica is one of the most important fish species being aquacultured in Japan but is hampered from the fact that this species does not reproduce in captivity. Artificial induction of gonadal maturation has been successful by administration of pituitary extracts or human chorionic gonadotropin, but the understanding the regulatory mechanism of gonadal development moderated by follicle stimulating hormone (FSH) and luteinizing hormone (LH) remains elusive due to lack of suitable amounts of eel gonadotropins (GTHs). In the present study, we produced tethered rec-eel LH and deglycosylated mutants (56, 79 and 56-79 of α subunit; 10 of β-subunit) of Asn-linked oligosaccharides in CHO suspension cells. Luteinizing hormone acts through binding to its specific receptor. Binding of ligand to the receptor activates the adenosine 3',5'-cyclic monophosphate (cAMP) pathway (McFarland et al., 1989; Ji and Ji, 1991a; Rose, 1998) and the inositol 1 phosphate (IP1) secondary messenger systems. After stimulation of eelLH/CG receptor transfected CHO cells with rec-LH wild type (wt) and mutant hormones as a ligand, production of cAMP and IP-1 were evaluated (Cisbio). cAMP IC-50 values by rec-eelLH wt; αΔ56; αΔ79; αΔ56_79 and βΔ10 were 606.2; 374.9; 100.3; 14.2 and 210.9 ng/ml, respectively. IP-1 IC-50 values by rec-eelLH wt; αΔ56; αΔ79; αΔ56.79 and βΔ10 were 28.3; 16.04; 4.3; 2.1 and 3.6ng/ml, respectively too. As seen in both of the second messenger production, general stimulatory pattern is analogous. cAMP and IP-1 stimulation by wild type and αΔ56, as well as αΔ79 and βΔ10 were approximate, but the stimulating effect of double mutant (αΔ56_ 79) was drastically higher. According to the data, deglycosylated eelLH may bind to the receptor with high affinity and cAMP production is gradually increased. Furthermore, receptor activation by tethered rec-eel mutant ligands (FreeStyle CHO-MAX Expression System) will be evaluated with β arrestin recruitment and GPCR internalization for N-linked oligosaccharides’ biological role in activation of eelLH/CGR.

국내에서도 소득증대와 시장 확대 등에 따라 기능성 식품에 대한 의존도가 증대되고 있다. 특히 유아용 및 노인용 식품제조에 적합한 유산균 제제의 개발과 함께 선발된 유산균의 생육을 위한 특정 생장촉진물질의 탐색이 필요한 실정이다. 따라서 본 연구에서는 국내 두유수유아의 분변으로부터 분리, 선발된 Bifidobacterium 균을 생균제로서의 능력을 향상시키기 위해 Bifidobacterium 균의 성장요인으로 알려져 있고, 인체에 유용한 기능성 올리고당의 첨가에 따른 생육특성을 조사하였다. 선발된 균주는 기능성 올리고당이 첨가된 배지에서 증식이 더 빠르게 나타났고, 그 중에서도 GOS가 5% 첨가된 배지에서 가장 빠르게 증식하였다. 또한, 기능성 올리고당을 첨가한 경우 α-galactosidase와 β-galactosidase의 활성이 더 좋아졌고, 균체의 세포막 지방산 조성을 비교해본 결과 Oleic acid (C18:1 ω9C) 함량 또한 월등히 높게 나타났다. pH 변화와 총산도 실험을 수행한 결과, 기능성 올리고당을 첨가하지 않은 배지에서 pH 5.79의 0.873%를 나타낸 반면, 5% GOS 가 첨가된 배지에서는 pH 3.6의 1.188%, 3% IMO는 pH 3.69의 1.134%, 그리고 FOS는 pH 3.71의 1.098%등으로, 현저히 낮은 pH를 나타냈다. 내담즙성의 실험결과 , 기능성 올리고당 중 5% GOS가 가장 높은 나타냈으나, 첨가하지 않은 경우보다는 낮은 수치를 나타냈다. 병원성균 성장저해 실험을 한 결과, 5% GOS를 첨가한 배지에서 자란 Bif. breve PBH-30이 E. coli O157, Salmonella typhimurium, Staphylococcus aureus 등과 같은 병원성 균을 더 효과적으로 저해하였으며, 특히 E. coli O157의 생육이 효과적으로 저해되었다. In vitro 부착능을 실험한 결과, 대부분의 Bif. breve PBH-30이 Caco-2 cell의 세포외막의 굴곡을 따라 부착되었음을 확인 하였다.

The purposes of this study were to investigate the Maillard reactions of some oligosaccharides with lysine and the antioxidative effects of the ethanol extracts from their reaction mixtures on the soybean oil. The Maillard reactions were carried out of 2% oligosaccharides such as palatinose (PN), fructooligosaccharide (FO), isomaltooligosaccharide (IMO) with 2% lysine (L) for 24 hours heating at 60, 80, 100℃. The color intensity of Maillard reaction mixtures were determined by UV-VIS spectrophotometer upon reaction time and temperature. And the antioxidative effects on the soybean oil of each ethanol extract from Maillard reaction mixture of each oligosaccharide were measured by peroxide value (POV). POV's of soybean oil including reaction extracts were determined regularly every 2 days during 20 days storaged at 60±1℃. The results were obtained as follows: 1. The color intensity of the Maillard reaction mixtures were raised highly as the browning temperature and time increased. The color intensity of PN L browning mixture was the highest. The order of high color intensity at 100℃ was PN L〉FO L〉Glu L〉IMO L. 2. Comparing the antioxidative effect of Maillard reaction product (at 100℃, for 12 hours) of each oligosaccharide to that of BHT and TBHQ, the order of high antioxidative effect was TBHQ〉IMO L〉BHT〉Glu L〉PN L〉FO L. 3. From these results, it was known that PN L shown as high brown color intensity was appeared low antioxidative effect, while IMO L shown as low brown color intensity was appeared high antioxidative effect. So, it was recognized that there was no relation between brown color intensity and antioxidative effect.

Oligossaccharides have beneficial effects on the immune system, atopy and skin health according to the prebiotic effect on intestinal microflora. Due to the excellent physiological functions of these oligosaccharides, the demand for prebiotics is rapidly increasing in the world. In this review, it has been shown that galactooligosaccharide( GOS) intake improves skin health and atopic eczema and fructooligosaccharide( FOS) prevent atopic dermatitis in human clinical trials. These results indicate that oligosaccharides are sufficiently applicable as a functional food for improving skin health and skin diseases.

The purpose of this study was to investigate the effects of envrionmental factors on the ecological responses, yield, and quality properties and physicochemical characteristics of yacon cultivated in various regions of Korea. Experiments were carried out in eight regions from 2010 to 2013. The temperature range in Jinbu, Bonghwa, Cheolwon, and Gangneung during the growth period of yacon cultivation was 17.5-24.6°C. The total yield and marketable yield of tuberous root in Jinbu were 4,065 and 3,196 kg/10a, respectively. The sugar content of yacon tuberous roots comprised 0.11-0.20% fructose, 0.11-0.37% glucose, 0.39-0.68% sucrose, 0.07-0.37% reducing sugar, and 7.03-9.62% fructo-oligosaccharides. The content of fructo-oligosaccharides, which is a functional substance, was the highest in yacon cultivated in Jinbu. Based on the productivity and functional fructo-oligosaccharides, the optimum areas to cultivate yacon are Jinbu and Bonghwa, which are located in the middle-highland zone (500-560 m) and have a suitable growth temperature of 18-25°C. The results of the present study indicate that temperature could be the factor with the greatest influence on the root growth of yacon.

Oligosaccharides have 2nd effects such as improvement of mineral absorption, immune system, atopy, skin health, and others, based on prebiotic effect. Because of outstanding physiological functions of oligosaccharides, the demand of prebiotics is dramatically increased together with probiotics in the world. Because oligosaccharides are distributed not only as general foods but as health functional foods, the demand of oligosaccharides especially in Korea can be expected to increase continuously. However, because each oligosaccharide has different stability in various temperature and pH conditions, it is very important to choose proper oligosaccharide considering heat and pH treatment in the processes, pH in finished products and storage temperatures, and pH condition. Meantime, because daily dosage of each oligosaccharide is different, it’s also important to choose ideal oligosaccharide considering types of foods and intake quantities.

This study was carried out to establish the analysis condition for oligosaccharides of soybeans (Glycine max L.) and to assay quantitative evaluation of oligosaccharide contents in soybean varieties and soybean sprouts. 55% acetonitrile solution used as a

대두에 함유되어 있는 Stachyose를 소화성당으로 변화시켜 식품으로서의 가치를 제고시킬 목적으로 대두 '평양'품종을 표준발아와 Hypochlorite를 처리한후 발아과정중 종자내에 함유되어있는 Monosaccharides 및 Oligosaccharides의 함량변화를 Thin-Layer Chromatography법으로 실험하였다. 1. Monosaccharides의 함량변화: 표준발아시 전기간중 Fructose가 제일함량이 많았고 Glucose, Galactose순으로 함량이 적었고 시간이 경과함에 따라 점점 감소되었다. Hypochlorite처리시 전기간중 Galactose는 전연 나타나지 않았다. 두 처리간에는 144시간을 제외하고 표준발아시켰을때가 Monosaccharide의 함량이 많았다. 2. Oligosaccharides의 함량변화: Sucrose는 표준발아시 함량은 적으나 전기간중 나타났으며96시간까지는 증가되었다가 약간씩 감소됐다.